Isolation and immunophenotyping by flow cytometry of canine peripheral blood and intraepithelial and lamina propria duodenal T lymphocytes

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dc.contributor.authorAgulla Pérez, Beatriz
dc.contributor.authorGarcía-Sancho Téllez, Mercedes Guadalupe
dc.contributor.authorSainz Rodríguez, Ángel
dc.contributor.authorRodríguez Franco, Fernando
dc.contributor.authorDíaz-Regañón Fernández, David Rafael
dc.contributor.authorRodríguez Bertos, Antonio Manuel
dc.contributor.authorVillaescusa Fernández, Alejandra
dc.date.accessioned2023-06-16T14:18:50Z
dc.date.available2023-06-16T14:18:50Z
dc.date.issued2021-07-28
dc.descriptionCRUE-CSIC (Acuerdos Transformativos 2021)
dc.description.abstractThe gut associated lymphoid tissue (GALT) effector sites play a crucial role on the pathogenesis of many immunemediated gastrointestinal diseases. The lymphocytes at these effector sites are principally T cells which present important morphological, phenotypical and functional differences. Flow cytometry (FC) is one of the most commonly used techniques to characterize intestinal lymphocytes in human and animal models. Published studies with a focus on dogs for intraepithelial lymphocytes (IEL) immunophenotyping exist in very limited numbers. Moreover, no lamina propria lymphocytes (LPL) isolation protocols in the canine species have been described for FC evaluation. In addition to immune intestinal dysregulation, imbalances in the peripheral blood immune system have been described in both human and animal gastrointestinal disorders. The aim of this study was to provide a protocol for canine IEL and LPL isolation for FC immunophenotyping of T cells subsets. Specifically, T helper, T cytotoxic, activated Th and Tc lymphocytes, regulatory, double negative, double positive, IFN-γ and IL-4 producing T cells, and to compare their respective populations between these effector sites and with the blood stream compartment in healthy dogs. The potential relationship of these cells distributions with age, sex and breed was also evaluated. This study included sixteen healthy dogs of different sexes and breeds with a mean age of 4.55 ± 2.93 years old. The selected protocols for the three immune compartments showed proper cell yield, purity, viability, and the absence of phenotypic and functional disturbances. Histologically, an adequate separation of the duodenal epithelium from the lamina propria was also observed. All the proposed T cells subsets were identified in the three immune compartments studied, showing some statistically significant differences in their distributions at these locations that highlight the importance of their individual evaluation. This study provides an adequate method for canine small intestine IEL and LPL isolation for FC immunophenotyping and is key for future studies on the gastrointestinal immune system associated with different canine diseases.
dc.description.departmentDepto. de Medicina y Cirugía Animal
dc.description.facultyFac. de Veterinaria
dc.description.facultyCentro de Vigilancia Sanitaria Veterinaria (VISAVET)
dc.description.refereedTRUE
dc.description.sponsorshipUniversidad Complutense de Madrid/Banco de Santander
dc.description.statuspub
dc.eprint.idhttps://eprints.ucm.es/id/eprint/69609
dc.identifier.doi10.1016/j.vetimm.2021.110305
dc.identifier.issn0165-2427
dc.identifier.officialurlhttps://doi.org/10.1016/j.vetimm.2021.110305
dc.identifier.urihttps://hdl.handle.net/20.500.14352/4641
dc.journal.titleVeterinary Immunology and Immunopathology
dc.language.isoeng
dc.page.initial110305
dc.publisherElsevier
dc.relation.projectID(PR41/17-21023)
dc.rightsAtribución-NoComercial-SinDerivadas 3.0 España
dc.rights.accessRightsopen access
dc.rights.urihttps://creativecommons.org/licenses/by-nc-nd/3.0/es/
dc.subject.keywordFlow cytometry
dc.subject.keywordIntraepithelial lymphocytes
dc.subject.keywordLamina propria lymphocytes
dc.subject.keywordT lymphocytes
dc.subject.keywordDog
dc.subject.keywordPeripheral blood lymphocytes
dc.subject.ucmVeterinaria
dc.subject.ucmBiología molecular (Biología)
dc.subject.unesco3109 Ciencias Veterinarias
dc.subject.unesco2415 Biología Molecular
dc.titleIsolation and immunophenotyping by flow cytometry of canine peripheral blood and intraepithelial and lamina propria duodenal T lymphocytes
dc.typejournal article
dc.volume.number239
dspace.entity.typePublication
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